LOCUS       sequence-349538-        5545 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  MoClo Level 1, position 2, transcriptional unit for transient 
            expression of major latex protein-like (NmMLPL) from Nepeta mussinii
            promoter regions contains 4x attB sites for recruitment fo 
            gal4AD-phiC31.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5545)
  AUTHORS   Dudley QM, Jo S, Guerrero DAS, Chhetry M, Smedley MA, Harwood WA, 
            Sherden NH, O'Connor SE, Caputi L, Patron NJ
  TITLE     Reconstitution of monoterpene indole alkaloid biosynthesis in genome
            engineered Nicotiana benthamiana.
  JOURNAL   Commun Biol. 2022 Sep 10;5(1):949. doi: 10.1038/s42003-022-03904-w.
  PUBMED    36088516
REFERENCE   2  (bases 1 to 5545)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Commun 
            Biol."; date: "2022-09-10"; volume: "5(1):949. doi"; pages: " 
            10.1038/s42003-022-03904-w"
COMMENT     Sequence Label: pEPQD1CB0792
FEATURES             Location/Qualifiers
     source          1..5545
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      28..659
                     /label=oriV
                     /note="incP origin of replication"
     misc_feature    751..775
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     protein_bind    892..925
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="minimal attB site for the phi-C31 integrase (Groth 
                     et al., 2000)"
     protein_bind    951..984
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="minimal attB site for the phi-C31 integrase (Groth 
                     et al., 2000)"
     protein_bind    1014..1047
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="minimal attB site for the phi-C31 integrase (Groth 
                     et al., 2000)"
     protein_bind    1073..1106
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="minimal attB site for the phi-C31 integrase (Groth 
                     et al., 2000)"
     promoter        1123..1169
                     /label=minimal CaMV 35S promoter
                     /note="minimal 35S promoter from cauliflower mosaic virus"
     misc_feature    1259..1314
                     /label=TMV Omega
                     /note="translational enhancer from the tobacco mosaic virus
                     5'-leader sequence (Gallie et al., 1988)"
     terminator      1800..2052
                     /label=MAS terminator
                     /note="mannopine synthase terminator"
     misc_feature    2098..2122
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             complement(2278..3426)
                     /codon_start=1
                     /product="trans-acting replication protein that binds to 
                     and activates oriV"
                     /label=trfA
                     /translation="MNRTFDRKAYRQELIDAGFSAEDAETIASRTVMRAPRETFQSVGS
                     MVQQATAKIERDSVQLAPPALPAPSAAVERSRRLEQEAAGLAKSMTIDTRGTMTTKKRK
                     TAGEDLAKQVSEAKQAALLKHTKQQIKEMQLSLFDIAPWPDTMRAMPNDTARSALFTTR
                     NKKIPREALQNKVIFHVNKDVKITYTGVELRADDDELVWQQVLEYAKRTPIGEPITFTF
                     YELCQDLGWSINGRYYTKAEECLSRLQATAMGFTSDRVGHLESVSLLHRFRVLDRGKKT
                     SRCQVLIDEEIVVLFAGDHYTKFIWEKYRKLSPTARRMFDYFSSHREPYPLKLETFRLM
                     CGSDSTRVKKWREQVGEACEELRGSGLVEHAWVNDDLVHCKR"
     rep_origin      complement(3762..4350)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3842..3861)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4521..5312,5313..5381))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     5144..5163
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5382..5486)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 cccctggggg gatgggaggc ccgcgttagc gggccgggag ggttcgagaa gggggggcac
       61 cccccttcgg cgtgcgcggt cacgcgcaca gggcgcagcc ctggttaaaa acaaggttta
      121 taaatattgg tttaaaagca ggttaaaaga caggttagcg gtggccgaaa aacgggcgga
      181 aacccttgca aatgctggat tttctgcctg tggacagccc ctcaaatgtc aataggtgcg
      241 cccctcatct gtcagcactc tgcccctcaa gtgtcaagga tcgcgcccct catctgtcag
      301 tagtcgcgcc cctcaagtgt caataccgca gggcacttat ccccaggctt gtccacatca
      361 tctgtgggaa actcgcgtaa aatcaggcgt tttcgccgat ttgcgaggct ggccagctcc
      421 acgtcgccgg ccgaaatcga gcctgcccct catctgtcaa cgccgcgccg ggtgagtcgg
      481 cccctcaagt gtcaacgtcc gcccctcatc tgtcagtgag ggccaagttt tccgcgaggt
      541 atccacaacg ccggcggccg cggtgtctcg cacacggctt cgacggcgtt tctggcgcgt
      601 ttgcagggcc atagacggcc gccagcccag cggcgagggc aaccagcccg gtgagcgtcg
      661 caaaggagat cctgatctga ctgatgggct gcctgtatcg agtggtgatt ttgtgccgag
      721 ctgccggtcg gggagctgtt ggctggctgg tggcaggata tattgtggtg taaacaaatt
      781 gacgcttaga caacttaata acacattgcg gacgttttta atgtactggg gtggatgcag
      841 tgggccccac tctgtgaaga caagcaagaa ttcaagcttg gagcggtgcg ggtgccaggg
      901 cgtgcccttg ggctccccgg gcgcgtactc cactccccga gccggtgcgg gtgccagggc
      961 gtgcccttgg gctccccggg cgcgtactcc actcccccca tggtgcggtg cgggtgccag
     1021 ggcgtgccct tgggctcccc gggcgcgtac tccactcccc gagccggtgc gggtgccagg
     1081 gcgtgccctt gggctccccg ggcgcgtact ccactcccct tcgcaagacc cttcctctat
     1141 ataaggaagt tcatttcatt tggagaggac tccggtattt ttacaacaat taccacaaca
     1201 aaacaaacaa caaacaacat tacaatttac tattctagtc gaatactgta tttttacaac
     1261 aattaccaac aacaacaaac aacaaacaac attacaatta ctatttacaa ttacaatggc
     1321 ttcgaagctt gaaatagaaa ttgagttgaa aagtgatgta gaaaagatgt ggaaacactt
     1381 caaagaattc accaaattat tccccaaggc ttgtcctcat ctttacgaga agattgatgt
     1441 gatcgaggga gatgggatat cagttggaac aatctttgtc agtaccctga aaccaacaga
     1501 gttgaacccc gtggttatgg tgacgaagga gaagattgat tttttagatg atgagaacaa
     1561 aatgttgagg tatagttaca tggaagggga gatattaaag aattacaaga atttgagggg
     1621 cacagtgcat atgagcagca gcaaaagtgg tggaactata ttcaaatatt caggagaatt
     1681 cgaaaaggca aatgaacaag tccctgatcc tgttttcttc aaagatttta tggtcatagt
     1741 tttccaaggt ttggatgatt atattctcaa gggcatgaac catacatgtc aaaattaagc
     1801 ttggactccc atgttggcaa aggcaaccaa acaaacaatg aatgatccgc tcctgcatat
     1861 ggggcggttt gagtatttca actgccattt gggctgaatt gtagacatgc tcctgtcaga
     1921 aattccgtga tcttactcaa tattcagtaa tctcggccaa tatcctaaat gtgcgtggct
     1981 ttatctgtct ttgtattgtt tcatcaattc atgtaacgtt tgcttttctt atgaattttc
     2041 aaataaatta tccgctacta ttgtcttctg cacgaagtgg tttaaactat cagtgtttga
     2101 caggatatat tggcgggtaa acctaagaga aaagagcgtt tattagaata atcggatatt
     2161 taaaagggcg tgaaaaggtt tatccgttcg tccatttgta tgtgcatgcc aaccacaggg
     2221 ttccccagat caggcgctgg ctgctgaacc cccagccgga actgacccca caaggcccta
     2281 gcgtttgcaa tgcaccaggt catcattgac ccaggcgtgt tccaccaggc cgctgcctcg
     2341 caactcttcg caggcttcgc cgacctgctc gcgccacttc ttcacgcggg tggaatccga
     2401 tccgcacatg aggcggaagg tttccagctt gagcgggtac ggctcccggt gcgagctgaa
     2461 atagtcgaac atccgtcggg ccgtcggcga cagcttgcgg tacttctccc atatgaattt
     2521 cgtgtagtgg tcgccagcaa acagcacgac gatttcctcg tcgatcagga cctggcaacg
     2581 ggacgttttc ttgccacggt ccaggacgcg gaagcggtgc agcagcgaca ccgattccag
     2641 gtgcccaacg cggtcggacg tgaagcccat cgccgtcgcc tgtaggcgcg acaggcattc
     2701 ctcggccttc gtgtaatacc ggccattgat cgaccagccc aggtcctggc aaagctcgta
     2761 gaacgtgaag gtgatcggct cgccgatagg ggtgcgcttc gcgtactcca acacctgctg
     2821 ccacaccagt tcgtcatcgt cggcccgcag ctcgacgccg gtgtaggtga tcttcacgtc
     2881 cttgttgacg tggaaaatga ccttgttttg cagcgcctcg cgcgggattt tcttgttgcg
     2941 cgtggtgaac agggcagagc gggccgtgtc gtttggcatc gctcgcatcg tgtccggcca
     3001 cggcgcaata tcgaacaagg aaagctgcat ttccttgatc tgctgcttcg tgtgtttcag
     3061 caacgcggcc tgcttggcct cgctgacctg ttttgccagg tcctcgccgg cggtttttcg
     3121 cttcttggtc gtcatagttc ctcgcgtgtc gatggtcatc gacttcgcca aacctgccgc
     3181 ctcctgttca agacgacgcg aacgctccac ggcggccgat ggcgcgggca gggcaggggg
     3241 agccagttgc acgctgtcgc gctcgatctt ggccgtagct tgctggacca tcgagccgac
     3301 ggactggaag gtttcgcggg gcgcacgcat gacggtgcgg cttgcgatgg tttcggcatc
     3361 ctcggcggaa aaccccgcgt cgatcagttc ttgcctgtat gccttccggt caaacgtccg
     3421 attcattcac cctccttgcg ggattgcccc gactcacgcc ggggcaatgt gcccttattc
     3481 ctgatttgac ccgcctggtg ccttggtgtc cagataatcc accttatcgg caatgaagtc
     3541 ggtcccgtag accgtctggc cgtccttctc gtacttggta ttccgaatct tgccctgcac
     3601 gaataccagc gaccccttgc ccaaatactt gccgtgggcc tcggcctgag agccaaaaca
     3661 cttgatgcgg aagaagtcgg tgcgctcctg cttgtcgccg gcatcgttgc gccacatcta
     3721 ggatctgcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     3781 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     3841 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     3901 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     3961 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     4021 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     4081 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     4141 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     4201 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     4261 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     4321 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     4381 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     4441 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     4501 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     4561 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     4621 gagggcttac catctggccc cagtgctgca atgataccgc gagaaccacg ctcaccggct
     4681 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     4741 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     4801 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
     4861 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     4921 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     4981 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     5041 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     5101 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
     5161 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     5221 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     5281 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     5341 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
     5401 tattgaagca tttatcaagg ttattgtctc atgagcggat acatatttga atgtatttag
     5461 aaaaataaac aaataggggt tccgcgcacg aattggccag cgctgccatt tttggggtga
     5521 ggccgttcgc ggccgagggg cgcag
//
